WITHDRAWN: AMPK α Succinylation modification of protein K266 enhances lipid metabolism and promotes growth of non-small cell lung cancer cells

Abstract

Objective: The succinylation level of the AMPKα1 protein at K266 is higher in non-small cell lung cancer tissues than normal tissues, but its relationship with NSCLC development is unclear and thus needs to be investigated. Methods: AMPKα1-knockout and mutant cell lines that simulate AMPKα1 succinylated and desuccinylated at K266 were generated via CRISPR/Cas9 and site-directed mutagenesis technology. The effects of AMPKα1 succinylation at K266 on the proliferation, apoptosis and lipid metabolism of NSCLC cells were also analysed. The role of succinyltransferases and desuccinylases in regulating AMPKα1 succinylation at K266 was analysed by immunoprecipitation and in vitro catalytic experiments. The effects of AMPKα1 succinylation on lipid metabolism and the AMPKα1 ubiquitination level were also analysed. The relationship between AMPKα1 succinylation at K266 and NSCLC development was analysed via immunohistochemistry. Results: The expression levelsof AMPKα1 and AMPKα1 succinylated at K266 were higher in NSCLC tissues, which promoted the growth of NSCLC. The 5-year overall survival rate of the group with high levels of AMPKα1 succinylated at K266 was lower than that of the group with low levels of succinylation at K266, and the succinylation levels were associated with the T (P=0.024), N (P=0.018), and TNM stages (P=0.024) in NSCLC patients. HAT1 and SIRT5 can interact with AMPKα1 and participate in the regulation of AMPKα1 succinylation at K266. The succinylation of AMPKα1 at K266 regulates lipid metabolism, and this process can lead to a decrease in AMPKα1 ubiquitination. Conclusion: AMPKα1 succinylation at K266 inhibits the ubiquitination-mediated degradation of AMPKα1, which increases the stability of this protein, enhances lipid metabolism, and promotes the growth of NSCLC cells. These findings have clinical diagnostic value for NSCLC and provide important clues for understanding the occurrence and development of NSCLC.